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Protein Engineering Strategies to Expand CRISPR-Cas9 Applications
Joint Authors
Ribeiro, Lucas F.
Ribeiro, Liliane F. C.
Barreto, Matheus Q.
Ward, Richard John
Source
International Journal of Genomics
Issue
Vol. 2018, Issue 2018 (31 Dec. 2018), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2018-08-02
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
The development of precise and modulated methods for customized manipulation of DNA is an important objective for the study and engineering of biological processes and is essential for the optimization of gene therapy, metabolic flux, and synthetic gene networks.
The clustered regularly interspaced short palindromic repeat- (CRISPR-) associated protein 9 is an RNA-guided site-specific DNA-binding complex that can be reprogrammed to specifically interact with a desired DNA sequence target.
CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as gene therapy, gene regulation, modifying epigenomes, and imaging chromosomes.
Although Cas9 has been successfully used as a precise tool in all these applications, some limitations have also been reported, for instance (i) a strict dependence on a protospacer-adjacent motif (PAM) sequence, (ii) aberrant off-target activity, (iii) the large size of Cas9 is problematic for CRISPR delivery, and (iv) lack of modulation of protein binding and endonuclease activity, which is crucial for precise spatiotemporal control of gene expression or genome editing.
These obstacles hinder the use of CRISPR for disease treatment and in wider biotechnological applications.
Protein-engineering approaches offer solutions to overcome the limitations of Cas9 and generate robust and efficient tools for customized DNA manipulation.
Here, recent protein-engineering approaches for expanding the versatility of the Streptococcus pyogenes Cas9 (SpCas9) is reviewed, with an emphasis on studies that improve or develop novel protein functions through domain fusion or splitting, rational design, and directed evolution.
American Psychological Association (APA)
Ribeiro, Lucas F.& Ribeiro, Liliane F. C.& Barreto, Matheus Q.& Ward, Richard John. 2018. Protein Engineering Strategies to Expand CRISPR-Cas9 Applications. International Journal of Genomics،Vol. 2018, no. 2018, pp.1-12.
https://search.emarefa.net/detail/BIM-1172679
Modern Language Association (MLA)
Ribeiro, Lucas F.…[et al.]. Protein Engineering Strategies to Expand CRISPR-Cas9 Applications. International Journal of Genomics No. 2018 (2018), pp.1-12.
https://search.emarefa.net/detail/BIM-1172679
American Medical Association (AMA)
Ribeiro, Lucas F.& Ribeiro, Liliane F. C.& Barreto, Matheus Q.& Ward, Richard John. Protein Engineering Strategies to Expand CRISPR-Cas9 Applications. International Journal of Genomics. 2018. Vol. 2018, no. 2018, pp.1-12.
https://search.emarefa.net/detail/BIM-1172679
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1172679